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Abstract

We introduce a method that combines two-photon polymerization (TPP) and surface functionalization to enable the indirect optical manipulation of live cells. TPP-made 3D microstructures were coated specifically with a multilayer of the protein streptavidin and non-specifically with IgG antibody using polyethylene glycol diamine as a linker molecule. Protein density on their surfaces was quantified for various coating methods. The streptavidin-coated structures were shown to attach to biotinated cells reproducibly. We performed basic indirect optical micromanipulation tasks with attached structure-cell couples using complex structures and a multi-focus optical trap. The use of such extended manipulators for indirect optical trapping ensures to keep a safe distance between the trapping beams and the sensitive cell and enables their 6 degrees of freedom actuation.

Figures (5)

Fig. 1 a) Drawing of microstructures polymerized onto a glass substrate for cell adhesion experiments: cross, ellipsoid and four-spheroid manipulator. b) Scheme of the indirect trapping of a cell with a four-spheroid manipulator. The four static trapping beams are arranged on the centers of the spheroids with holographic optical tweezers.

Fig. 4 Indirect optical manipulation of K562 cells (a-f) showing the sequence of the trapping and dragging of a cell with a streptavidin-coated cross (a) (see Visualization 1). The trapped cross is rotated around the optical axis by changing the polarization direction of the trapping beam (b, c). The trapped cross attaches to the cell (d). The cell is dragged around the reference cell marked with asterisk (e, f). Position distribution (g) of the center of an optically trapped ellipsoid dragging a cell; insert: the image of the ellipsoid-cell couple. Crosshairs indicate trap position.

Fig. 5 Translation of an indirectly trapped K562 cell in its medium with a four-spheroid manipulator along the three coordinate axes: (a-c) dragging along the x axis (see Visualization 2), (e)-(f) along the y axis (see Visualization 3) and (h)-(j) along the z (optical) axis. The histograms on panel (d) show the displacement of the two spheroids marked with asterisk during x-drag and that on (g) shows the tilt of the entire structure during y-drag. The two lines on panel (e) indicate the maximum tilt angle. Scale bar: 10 μm.